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Characterization of Multi-Functional Properties and Conformational Analysis of MutS2 from Thermotoga maritima MSB8
The MutS2 homologues have received attention because of their unusual activities that differ from those of MutS. In this work, we report on the functional characteristics and conformational diversities of Thermotoga maritima MutS2 (TmMutS2). Various biochemical features of the protein were demonstra...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2012
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3335848/ https://www.ncbi.nlm.nih.gov/pubmed/22545085 http://dx.doi.org/10.1371/journal.pone.0034529 |
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author | Jeong, Euiyoung Jo, Hunho Kim, Tae Gyun Ban, Changill |
author_facet | Jeong, Euiyoung Jo, Hunho Kim, Tae Gyun Ban, Changill |
author_sort | Jeong, Euiyoung |
collection | PubMed |
description | The MutS2 homologues have received attention because of their unusual activities that differ from those of MutS. In this work, we report on the functional characteristics and conformational diversities of Thermotoga maritima MutS2 (TmMutS2). Various biochemical features of the protein were demonstrated via diverse techniques such as scanning probe microscopy (SPM), ATPase assays, analytical ultracentrifugation, DNA binding assays, size chromatography, and limited proteolytic analysis. Dimeric TmMutS2 showed the temperature-dependent ATPase activity. The non-specific nicking endonuclease activities of TmMutS2 were inactivated in the presence of nonhydrolytic ATP (ADPnP) and enhanced by the addition of TmMutL. In addition, TmMutS2 suppressed the TmRecA-mediated DNA strand exchange reaction in a TmMutL-dependent manner. We also demonstrated that small-angle X-ray scattering (SAXS) analysis of dimeric TmMutS2 exhibited nucleotide- and DNA-dependent conformational transitions. Particularly, TmMutS2-ADPnP showed the most compressed form rather than apo-TmMutS2 and the TmMutS2-ADP complex, in accordance with the results of biochemical assays. In the case of the DNA-binding complexes, the stretched conformation appeared in the TmMutS2-four-way junction (FWJ)-DNA complex. Convergences of biochemical- and SAXS analysis provided abundant information for TmMutS2 and clarified ambiguous experimental results. |
format | Online Article Text |
id | pubmed-3335848 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-33358482012-04-27 Characterization of Multi-Functional Properties and Conformational Analysis of MutS2 from Thermotoga maritima MSB8 Jeong, Euiyoung Jo, Hunho Kim, Tae Gyun Ban, Changill PLoS One Research Article The MutS2 homologues have received attention because of their unusual activities that differ from those of MutS. In this work, we report on the functional characteristics and conformational diversities of Thermotoga maritima MutS2 (TmMutS2). Various biochemical features of the protein were demonstrated via diverse techniques such as scanning probe microscopy (SPM), ATPase assays, analytical ultracentrifugation, DNA binding assays, size chromatography, and limited proteolytic analysis. Dimeric TmMutS2 showed the temperature-dependent ATPase activity. The non-specific nicking endonuclease activities of TmMutS2 were inactivated in the presence of nonhydrolytic ATP (ADPnP) and enhanced by the addition of TmMutL. In addition, TmMutS2 suppressed the TmRecA-mediated DNA strand exchange reaction in a TmMutL-dependent manner. We also demonstrated that small-angle X-ray scattering (SAXS) analysis of dimeric TmMutS2 exhibited nucleotide- and DNA-dependent conformational transitions. Particularly, TmMutS2-ADPnP showed the most compressed form rather than apo-TmMutS2 and the TmMutS2-ADP complex, in accordance with the results of biochemical assays. In the case of the DNA-binding complexes, the stretched conformation appeared in the TmMutS2-four-way junction (FWJ)-DNA complex. Convergences of biochemical- and SAXS analysis provided abundant information for TmMutS2 and clarified ambiguous experimental results. Public Library of Science 2012-04-24 /pmc/articles/PMC3335848/ /pubmed/22545085 http://dx.doi.org/10.1371/journal.pone.0034529 Text en Jeong et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Jeong, Euiyoung Jo, Hunho Kim, Tae Gyun Ban, Changill Characterization of Multi-Functional Properties and Conformational Analysis of MutS2 from Thermotoga maritima MSB8 |
title | Characterization of Multi-Functional Properties and Conformational Analysis of MutS2 from Thermotoga maritima MSB8 |
title_full | Characterization of Multi-Functional Properties and Conformational Analysis of MutS2 from Thermotoga maritima MSB8 |
title_fullStr | Characterization of Multi-Functional Properties and Conformational Analysis of MutS2 from Thermotoga maritima MSB8 |
title_full_unstemmed | Characterization of Multi-Functional Properties and Conformational Analysis of MutS2 from Thermotoga maritima MSB8 |
title_short | Characterization of Multi-Functional Properties and Conformational Analysis of MutS2 from Thermotoga maritima MSB8 |
title_sort | characterization of multi-functional properties and conformational analysis of muts2 from thermotoga maritima msb8 |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3335848/ https://www.ncbi.nlm.nih.gov/pubmed/22545085 http://dx.doi.org/10.1371/journal.pone.0034529 |
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